Dynamics of dendritic cell‐derived vesicles: high‐resolution flow cytometric analysis of extracellular vesicle quantity and quality
[摘要] Nano‐sizedmembranevesiclesaresecretedbymanycelltypes.Thesevesiclescanserveascarriersofcellularinformation.DC‐derivedvesiclescanbetargetedtootherimmunecellsandmodifytheirfunction.AccurateanalysisofquantitativeandqualitativechangesinEVproductionbyDCupondifferentactivationstimuliisneededtofurtherrevealtheimmuneregulatorypropertiesofDC‐derivedEVs.However,methodsforreliablequantificationofindividualEVsandforanalysisoftheheterogeneityofEVpopulationsarelimited.Withourrecentlydevelopedhigh‐resolutionflowcytometry‐basedmethod,wecanperformahigh‐throughput,multiparameter,andquantitativeanalysisofindividualEVs.Withtheuseofthisnoveltechnique,weshowthatdespitepreviousassumptions,stimulationwithbacterialLPSincreasesEVreleasebyDC.Furthermore,wedemonstrateheterogeneityinDC‐derivedEVsregardingtheirbuoyantdensityandMHCclassIIcontent.Finally,weshowthatcognateinteractionbetweenLPS‐stimulatedDCandCD4+TcellsaffectsboththequantityandqualityofLPSDC‐derivedEVspresentintheculturesupernatant.Thesedataindicatethatflowcytometry‐basedanalysisofindividualEVsisavaluable,noveltooltostudythedynamicsofEVsecretionandcomposition,offeringgreatopportunitiestounveilthefunctionofimmunecell‐derivedEVs...
[发布日期] [发布机构]
[效力级别] [学科分类] 生理学
[关键词] exosomes;microvesicles;immune regulation;single vesicle analysis;cognate interaction;MHC class II [时效性]
